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Publication Number
US-8224058-B2
Publication Date
2012-07-17
Expiration Date
Abstract
A measurement apparatus for enumeration of particles or white blood cells in a sample comprises: a holder, which is arranged to receive a sample acquiring device that holds a sample, an imaging system, comprising a magnifying means and at least one digital image acquiring means, said imaging system being arranged to acquire at least one digital image of the sample, and an image analyser, which is arranged to analyse the digital image for identifying particles or white blood cells and determining the number of particles or white blood cells and which is arranged to analyse the digital image for identifying particles or white blood cells that are imaged in focus, determining types of these particles or white blood cells, the types being distinguished by physical features and determining the ratio of different types of particles or white blood cells.
Core Innovation
The invention relates to a measurement apparatus for enumeration of white blood cells in a sample. The apparatus includes a holder arranged to receive a sample acquiring device with a measurement cavity that holds a stained and hemolysed sample, and the measurement cavity is adapted for a uniform depth of 50-200 micrometers.
The apparatus includes an imaging system adapted to acquire a plurality of digital images of the sample at different levels in a direction of depth of field using different optical settings. The imaging system is arranged to obtain the plurality of digital images with a depth of field in the range of 2-30 micrometers, supporting identification of stained white blood cells, determination of the number of white blood cells, and identification of white blood cells imaged in focus for type determination.
An image analyzer determines whether each white blood cell in an image is imaged in focus by using a value of a property falling within a predetermined range. For a specific white blood cell, the number of images in which the white blood cell is imaged is determined by counting from an image in which the white blood cell is determined to be out of focus in a first direction to an image in which the white blood cell is determined to be out of focus in a second direction, and based at least in part on the number of images, the image analyzer determines types and number of white blood cells imaged in focus and thereby determines the ratio of different types.
Claims Coverage
The partial content includes three independent claim sets: a measurement apparatus for enumeration, a method for enumeration, and computer-readable media implementing image acquisition and analysis. Across these independent claims, the inventive features consistently include depth-defined measurement, plurality of depth-of-field images acquired using different optical settings, in-focus determination based on a property within a predetermined range, and type and count determination based at least in part on the number of in-focus images for each white blood cell.
Depth-defined measurement cavity with depth-limited imaging slices
A measurement apparatus comprising a holder arranged to receive a sample acquiring device having a measurement cavity holding a stained and hemolysed sample with a uniform depth of 50-200 micrometers, and an imaging system acquiring a plurality of digital images at different levels in a direction of depth of field using different optical settings, with a depth of field range of 2-30 micrometers.
Focus-based identification and in-focus counting across depth directions
An image analyzer arranged to determine for each stained white blood cell whether it is imaged in focus when a value of a property falls within a predetermined range, and to determine the number of images in which a specific white blood cell is imaged by counting from an out-of-focus image in a first direction to an out-of-focus image in a second direction.
Type determination using physical features of in-focus cells and ratio based on image counts
An image analyzer arranged to identify stained white blood cells imaged in focus and determine types and number of the white blood cells imaged in focus, wherein types are distinguished by physical features of the stained white blood cells, whereby the ratio of different types of white blood cells in the sample is determined based at least in part on the number of images.
Enumeration method with reagent mixing and plurality of depth-of-field images
A method for enumeration comprising acquiring a sample into a measurement cavity of a sample acquiring device mixed with a reagent including a hemolysing agent for lysing red blood cells and a staining agent for staining white blood cells, wherein the measurement cavity has a uniform depth of 50-200 micrometers, acquiring a plurality of digital images of an irradiated sample at different levels using different optical settings with a depth of field in the range of 2-30 micrometers, and digitally analyzing each image for identifying stained white blood cells and determining the number.
Method-based focus criterion and counting of in-focus images for typing
Digitally analyzing each acquired digital image for identifying stained white blood cells imaged in focus and determining types and number of white blood cells imaged in focus, determining a white blood cell to be in focus when a value of a property falls within a predetermined range, and determining for a specific white blood cell the number of images in which it is imaged by counting from an out-of-focus image in a first direction to an out-of-focus image in a second direction, wherein determining the types and number is based at least in part on the number of images.
Non-transitory medium executing an enumeration method with in-focus image counting
A non-transitory computer-readable medium storing a computer program product causing a computer to execute a method comprising digitally analyzing a plurality of images to determine a number of white blood cells and identifying types in a focused region, where the in-focus determination uses a property value within a predetermined range, where for a specific white blood cell the number of images in which it is imaged is determined by counting from out-of-focus in a first direction to out-of-focus in a second direction, and where determining types and number of white blood cells imaged in focus is based at least in part on the number of images.
Non-transitory medium executing analysis to determine counts, types, and ratios
A non-transitory computer-readable medium storing a computer program comprising analyzing a plurality of digital images for identifying white blood cells and determining the number, acquiring the plurality of images of an irradiated sample at different depth levels using different optical settings with a depth of field in the range of 2-30 micrometers in a measurement cavity of uniform depth 50-200 micrometers, analyzing images for identifying white blood cells imaged in focus and determining types and number of white blood cells imaged in focus with types distinguished by physical features, determining a white blood cell in focus when a property value falls within a predetermined range, counting in-focus images for a specific white blood cell from out-of-focus in a first direction to out-of-focus in a second direction, and determining types and number based at least in part on the number of images.
Across the independent claims, the core coverage is for white blood cell enumeration and typing using a depth-defined measurement cavity of 50-200 micrometers, acquisition of a plurality of depth-of-field-limited digital images with different optical settings, determining in-focus cells using a property within a predetermined range, and determining types and counts, and thus ratios, based at least in part on the number of images in which each white blood cell is imaged in focus.
Stated Advantages
Documented Applications
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